Inhibitors of lipoxygenase metabolism exert synergistic effects with retinoic acid on differentiation of human leukemia HL-60 cells

Abstract

The interaction between drugs suppressing the production of arachidonic acid metabolites and inducers of granulocytic differentiation, i.e., all- trans retinoic acid and dimethyl sulphoxide (DMSO) was investigated using the human myeloid leukemia HL-60 cell line. The experiments were designed as a complete factorial combination of treatments and used chemiluminescence as a marker of cell oxidative burst (level of differentiation). It was clearly demonstrated that two structurally different inhibitors of 5-lipoxygenase metabolism, i.e., 3-[1-(4-chlorobenzyl)-3- t-butyl-thio-5-isopropylindol-2-yl]-2,2-dimethyl propanoic acid (MK-886) and esculetin, significantly potentiated the HL-60 cell differentiation induced by retinoic acid or DMSO. Detailed mathematical evaluation of the results revealed the synergistic character of the interaction. The most significant effects were achieved with a combination of 5-lipoxygenase inhibitors and low doses of retinoic acid. These results were confirmed by analysis of cell morphology and expression of cell surface antigen CD 11b after treatment of the cells with selected concentrations of agents. In contrast to those on differentiation, no additional effects of MK-886 or esculetin on cell proliferation (cell number and cell cycle parameters) and apoptosis were observed. An inhibitor of cyclooxygenases, indomethacin, affected neither cell proliferation nor differentiation of cells. The results implied that either modulation of 5-lipoxygenase metabolism or a certain type of imbalance in arachidonic acid metabolism could modulate the effects of retinoic acid or DMSO on myeloid cell differentiation.